【作者】 王玲；

【导师】 屈凌波； 李建军；

【作者基本信息】 郑州大学 ， 分析化学， 2006， 硕士

【摘要】 黄酮类化合物是一类存在于多种植物中的多酚化合物，研究发现它们具有广泛的药理活性，如抗氧化、抗肿瘤、抗病毒、抗炎等活性。蛋白质是重要的生命物质，在生物体内具有重要生理功能。大量生物化学和分子生物学调查研究表明，蛋白质是具有药理活性的黄酮类天然产物的靶分子。药效小分子与蛋白相互作用的研究对新型药物的设计、开发以及促进信息科学的发展都有重要意义。 本文主要是以分子光谱方法(荧光、紫外-可见光谱)对不同结构的黄酮、异黄酮及其磷酰化产物与蛋白的相互作用情况进行研究。通过对比研究结果，试图找到黄酮类天然产物与蛋白结合作用的规律。所获结果可为更深入地了解黄酮与蛋白的作用机理、研究黄酮分子的结构与蛋白作用关系提供有益的参考。 本文主要内容如下： 1．利用荧光光谱法研究了黄酮(芹菜素、木犀草素、槲皮素、芦丁)在生理pH下与牛血清白蛋白的相互作用。结果显示：在一定浓度范围内它们均能和蛋白发生弱相互作用；黄酮与蛋白的结合能力不仅与羟基个数有关，而且与羟基的位置也关系重大；B环3′，4′羟基邻位可以大大增强黄酮与牛血清白蛋白的结合能力；C环3位羟基是一很特殊的作用位点，不利于黄酮与蛋白的相互作用；药效分子体积过大造成的较大的空间位阻也不利于黄酮小分子和蛋白的结合。 2．应用荧光光谱法研究了异黄酮(染料木素、葛根素、大豆甙)与牛血清白蛋白的弱相互作用。结果表明：A环5位羟基可以增强异黄酮与蛋白的结合能力，可能是药效基团；葡萄糖(Glc)基的取代，有利于异黄酮与蛋白的结合。 3．以葛根素为母体，利用Atherton-Todd反应得到了葛根素的两个磷酸化产物(7-二乙基磷酸葛根素酯和7-二异丙基磷酸葛根素酯)；以荧光光谱法研究了葛根素及其磷酰化产物与牛血清白蛋白弱相互作用。结果显示葛根素及其磷酰化产物均能与BSA发生相互作用，磷酰化产物与蛋白的结合作用相对较弱。由实验结果推断A环7位羟基可能是影响黄酮与蛋白作用的重要基团，且当分子空间体积增大到一定程度时，位阻效应成为影响小分子与蛋白结合的重要因素。 4．分别从牛脑牛心中提取了钙调素(CaM)和磷酸二酯酶I(PDEI)；利用Atherton-Todd反应合成了5.7-二羟基磷酰化黄酮。并进一步研究了5.7-二羟基黄酮及更多还原

【Abstract】 The research discoveries that flavones compounds have many potential pharmaceutical usages, such as their anti-oxidation,anti-virus and anti-tumor effects. The proteins are important chemical substance in our life. A number of biochemical and molecular biological investigations have revealed that proteins are frequently the targets for therapeutically active flavonoids of both natural and synthetic origin. More scientists pay attention to the study of the interaction of small molecular and protein, which are benefit to design the new factionlized medicine to surmount many difficult diseases and to development of information science.This paper is mainly to study the interactions of different structural flavones, isflavones and their phosphorylated products with protein under physiological pH by means of fluorescence and UV-Vis spectroscopy. The relationship between the structure characteristics of these compounds and binding properties of the flavonoids and protein was preliminarily discussed. The knowledge obtained of the structure features that determine the binding capacity of these compounds and protein may make us better understand the mechanics of action and pharmacological activities of them and open up new avenues for the design of the most suitable flavonoid derivatives with structural variants.The main content of this thesis is as follows:First, the interactions of flavonoids, including luteolin, apigenin, quercetin and rutin, with bovine serum albumin(BSA) were studied under physiological conditions by fluorescence spectroscopic technique. The results showed that these compounds all could form non-covalent complexes with BSA at some range of the concentration, also the number and situation of hydroxyl of the flavonoids had an important influence on the binding potency towards protein. B(3’)-OH,B(4’)-OH strengthened the interaction of flavonoids and BSA, but the function of C(3)-OH was reversed relatively. Besides, the interaction between flavonoids and BSA can occur only when these compounds near BSA to some extent. The extent that small molecule close to biomolecule is relate to its tridimensional volume, taking this into account, the size of更多还原